PowerTaq 2X PCR Mix with Dye V2
Product Components
| Components | Component Number | 1 mL | 5 mL | 100 mL |
|---|---|---|---|---|
| PowerTaq 2X PCR Mix with Dye V2 | MZ20395 | 1 mL | 1 mL × 5 | 1 mL × 100 |
Product Description
PowerTaq 2X PCR Mix with Dye V2 is an optimized premix designed for fast PCR amplification, with extension rates of up to 10 s/kb, helping to significantly reduce PCR reaction time. The reagent includes DNA polymerase, dNTPs, MgCl₂, KCl, and other stabilizers. Only primers and template DNA need to be added to perform amplification.
This product contains a loading buffer, allowing PCR products to be directly loaded onto agarose gels for electrophoresis without adding additional loading dye.
PowerTaq 2X PCR Mix with Dye V2 can amplify up to a 5 kb target fragment from complex genomic DNA or up to a 10 kb target fragment from simple templates such as lambda DNA. It is suitable for Colony PCR, genotype identification, vector construction, and other routine PCR applications.
5′–3′ Exonuclease Activity
No
3′–5′ Exonuclease Activity
Yes
Fidelity
6X Taq
Product End
Blunt end
Storage
−20°C
Operation Description
Standard Protocol
It is recommended to prepare all reaction components on ice and then quickly transfer the reaction mixture to a thermocycler preheated to 98°C.
Recommended Reaction
| Component | 25 µL Reaction | 50 µL Reaction | Final Concentration |
|---|---|---|---|
| PowerTaq 2X PCR Mix with Dye V2 | 12.5 µL | 25 µL | 1X |
| Forward Primer (10 µM) | 0.5 µL | 1 µL | 0.2 µM |
| Reverse Primer (10 µM) | 0.5 µL | 1 µL | 0.2 µM |
| DNA Template* | Variable | Variable | <300 ng |
| Nuclease-free Water | To 25 µL | To 50 µL | N/A |
Note: The optimal reaction concentration varies depending on different DNA templates.
For a 50 µL reaction, the recommended DNA template amounts are provided separately.
For long-fragment amplification, the amount of template input can be appropriately increased.
| DNA Type | Input Amount |
|---|---|
| Plant, animal, and human genomic DNA | 10–100 ng |
| E. coli, lambda genomic DNA | 10 pg–200 ng |
| Plasmid DNA | 1 pg–10 ng |
| cDNA | 1–5 µL (less than 10% of total reaction volume) |
Recommended PCR Program
| Step | Temperature | Time | Cycles |
|---|---|---|---|
| Initial Denaturation | 98°C | 45 s* | 1 |
| Denaturation | 98°C | 10 s | 30 |
| Annealing | 55–65°C** | 30 s | 30 |
| Extension | 72°C | 10–30 s/kb*** | 30 |
| Final Extension | 72°C | 5 min | 1 |
| Hold | 4–12°C | — | 1 |
Notes
* For high-complexity DNA templates (e.g., high GC content), extend the initial denaturation time to 3 minutes.
For colony PCR, extend to 8 minutes to ensure complete denaturation.
** The annealing temperature can generally be set to 60°C by default.
For primers longer than 20 nt: use (lower primer Tm + 3°C)
For primers shorter than 20 nt: use the lower primer Tm
***
For low-complexity amplicons (e.g., plasmid DNA, lambda DNA) or fragments ≤1 kb: use 10 s/kb
For high-complexity amplicons or fragments >1 kb: increase extension time to 20–30 s/kb